The level of ROS was measured by flow cytometry and confocal microscopy. For the flow cytometry analysis, cells were incubated with MitoSOX (2 µM, Thermo Fisher Scientific, Sunnyvale, CA, USA) for 30 min, washed twice with PBS and analyzed in a flow cytometer (Beckman, USA). For the fluorescence microscopy assay, HK2 cells were incubated with DCFH-DA (10 μM, Beyotime Biotechnology, Shanghai, China) or MitoSOX (5 µM) for 30 min at 37 °C. The stained cells were then washed with PBS and observed using a fluorescence microscope (Olympus, Japan).

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